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The plasmid is now considered recombinant as it contains the human gene inserted into its DNA sequence. This engineered plasmid can be used in genetic engineering techniques to produce proteins or study gene function.
to make bacteria take up a plasmid, you need to add Ca+2 into solution to make the membrane porous. also to select for the bacteria that has the plasmid you need to have something like an ampicilin resistance sequence in there as well and grow the culture in an amp culture
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In the process of human gene cloning using recombinant plasmids what is the bacterial plasmid?
The bacterial plasmid is a small circular DNA molecule that is used as a vector to carry the gene of interest in gene cloning experiments. It is introduced into bacteria, where it replicates independently from the bacterial chromosome. The gene of interest is inserted into the plasmid using restriction enzymes and ligase.
How is a human gene recombined into a bacterial plasmid?
A human gene can be recombined into a bacterial plasmid using restriction enzymes to cut both the gene and the plasmid at specific sites. The cut gene can then be ligated into the cut plasmid using DNA ligase. The recombinant plasmid can then be introduced into bacteria through a process like transformation, allowing the bacteria to express the human gene.
What are the genes on the pVIB plasmid?
The pVIB plasmid contains various genes that confer antibiotic resistance and help in plasmid replication and maintenance. Some common genes found on pVIB include those encoding for beta-lactamase enzymes, which provide resistance to beta-lactam antibiotics, and replication proteins that ensure the plasmid is replicated and passed on to daughter cells during cell division.
Which enzyme do scientists use to bond a new gene to plasmid DNA?
Scientists use DNA ligase to bond a new gene to plasmid DNA. DNA ligase catalyzes the formation of phosphodiester bonds between the ends of the new gene and the plasmid, creating a recombinant DNA molecule.
Bacteria containing a plasmid into which the eukaryotic gene has integrated would grow where?
The bacteria containing the plasmid with the integrated eukaryotic gene would grow in a selective medium that supports the growth of bacteria carrying the plasmid. This medium would typically contain an antibiotic or a specific nutrient that selects for bacteria with the plasmid.
A plasmid that contains a gene for human growth hormone is and example of what?
A plasmid containing a gene for human growth hormone can be used in genetic engineering to produce recombinant human growth hormone. This plasmid can be introduced into host cells, such as bacteria, for the production of the hormone on a large scale.
What is the term for a plasmid that contains a foreign gene?
Recombiant DNA
How does a human insulin genes become part of a plasmid?
Human insulin genes can be inserted into a plasmid using recombinant DNA technology. This involves isolating the insulin gene from human cells, cutting the plasmid with a restriction enzyme, and then ligating the insulin gene into the plasmid. The plasmid can then be introduced into bacterial cells for replication and production of insulin.
Why does a gene fit into the opening in the plasmid?
The gene fits into the opening in the plasmid because the ends of the gene and the plasmid have been cut by specific enzymes to create complementary "sticky ends" that can bind together. This process is known as ligation, which joins the gene and the plasmid together to create a recombinant DNA molecule.
Why does bacteria that contains plasmid glow in uv light?
the plasmid contains a certain gene, which codes for the "Green Flourescent Protein." So you put the plasmid in the bacteria, the plasmid starts making that protein in the bacteria, and boom you've got glowing bacteria. works for bunnies and monkeys too, apparently =)
In the process of human gene cloning using recombinant plasmids what is the bacterial plasmid?
The bacterial plasmid is a small circular DNA molecule that is used as a vector to carry the gene of interest in gene cloning experiments. It is introduced into bacteria, where it replicates independently from the bacterial chromosome. The gene of interest is inserted into the plasmid using restriction enzymes and ligase.
When is a plasmid considered a recombinant plasmid?
A plasmid is considered recombinant when it contains DNA sequences from two different sources that have been artificially combined, often through genetic engineering techniques like restriction enzyme digestion and ligation. This results in a plasmid with modified or additional genetic material compared to its original form.
Why is your plasmid considered recombinant DN?
A plasmid is considered recombinant DNA when it contains DNA sequences from multiple sources that have been artificially joined together using molecular cloning techniques. This can include the insertion of a gene of interest into the plasmid for expression in a host organism, or the addition of regulatory elements to control gene expression.
How do you express a human gene in E. coli?
E.coli is used to express human genes because it can be easily grown in the lab. The gene is extracted from the DNA (by doing a partial digest with a restriction enzyme), and given a cohesive sticky end with a linker or adapter. It is then ligated to a plasmid vector, which had a restriction site compatible with the ends on the gene, eg if the plasmid contains a BamH1 site then you would add a linker or adapter which is compatible with the 5'GGATCC3' BamH1 recognition sequence. The cells are transformed (made to take up the plasmid vector) by chemical treatment; they are mixed with the plasmid, then a strong concentration of calcium (Ca2+) ions is added to the mixture to make the E.coli's membranes porous. The mixture is then heated to heat-shock the cells, to approx 50 degrees C for one minute. They are then cooled and allowed to recover in a nutrient rich broth at optimum temperature. This is a very inefficient process - only about 1 cell in every million is transformed. The pUC18 plasmid vector is useful because it contains the gene for ampicillin resistance. Any cells which subsequently grow on a medium containing ampicillin, therefore, have been transformed with the plasmid vector. It is also useful because it contains a beta-galactosidase gene, which itself contains the recognition site for a number of restriction enzymes, including BamH1. This is good because you can tell if the vector has taken up the gene you are trying to express when the vector no longer codes for the beta-galactosidase protein product. If the vector has been ligated with the gene, the gene will have disrupted the beta-galactosidase gene. This can be tested with IPTG (an auto-inducer) and X-gal, which will turn colonies of E.coli with the beta-galactosidase gene intact blue (ie, those without the gene of interest). Colonies which have had their beta-galactosidase gene destroyed by the ligation of the gene of interest will be colourless in the presence of X-gal and IPTG. These colonies are all clonal, so all cells in colourless colonies contain copies of the pUC18 plasmid vector which has been ligated with the human gene.
Can genetic engineering techniques treat cystic fibrosis?
Yes, there are two similar techniques in which i am aware of.AdenovirusesFirstly adenoviruses are made harmless by interefering with a gene involved in replication. A healthy from of the CFTR gene is extracted and cut with restriction endonucleases, the same enzyme is used to cut a bacterial plasmid. The gene and plasmid are mixed together along with DNA ligase to anneal the phosphosugar framework of the DNA fragment and bacterial plasmid. The plasmid is then mixed with epithelial cells. The plasmid is then isolated and purified and places into adenoviruses. These are then placed onto the nostrils of individuals with cystic fibrosis. The viruses find their way to epithelial cells in the airways and injected their DNA. The DNA contains the functional CFTR gene, the cells can then produce fucntional CFTR proteins.LiposomesA healthy gene is extracted from a human. This gene is then inserted into a bacterial plasmid, in a similar manner as discussed above. The bacterial plasmids are then inserted into bacteria. These are allowed to grow and divide, producing large quantities of the plasmid, with the required gene. These plasmids are then extracted and coated in a lipid soluble substance. They are then put into nasal sprays and sprayed onto the nostrils of effected individuals.
What type of gene is used to distinguish bacteria that carry a plasmid containing foreign DNA from those that don' t?
The plasmid that contains foreign DNA is engineered to also carry an antibiotic resistance gene. This antibiotic resistance gene codes for a protein that is able to inactivate an antibiotic thus keeping the cell alive. In the absence of the antibiotic resistance gene, the cells would not survive when exposed to an antibiotic. After transfection (the process of inserting the plasmid carrying the foreign gene into cells), the cells are gown in media containing an antibiotic. Cells that contain the plasmid (and therefore contain the antibiotic resistance gene) are able to survive in this medium. Cells that do not contain the plasmid (and therefore lack the antibiotic resistance gene) do not survive in this medium. The process described above is called selection
What is an extra loop of DNA that contains antibiotic resistance what gene?
An extra loop of DNA that carries antibiotic resistance genes is called a plasmid. These genes can provide bacteria with the ability to survive exposure to antibiotics.
